1. Field
The present disclosure relates to shuttle vectors for Corynebacterium and Escherichia coli, and promoter screening and gene product production using the same.
2. Description of the Related Art
Corynebacterium is a gram-positive bacterial strain widely used for the production of amino acids, such as glutamate, lysine, and threonine, and purine based nucleic acids, such as inosinic acid. Corynebacterium glutamicum, in particular, is easy to grow, tolerates high concentration cultivation (e.g., cultivation at concentrations up to approximately four times greater than concentrations tolerated by Escherichia coli (E. coli)), and has a stable genome that resists mutations. In addition, Corynebacterium glutamicum has several merits as an industrial strain, such as being a nonpathogenic strain, not forming spores, thus having no deleterious effects on the environment, and the like.
A cloning vector is a cyclic DNA which can be replicated independently from a main chromosome in bacteria. A cloning vector includes an origin of replication for maintenance of a plasmid form within the strain, a selectable marker gene for selection of strains having the cloning vector, and a multi-cloning site (MCS) for cloning an exogenous gene.
A shuttle vector generally includes a vector which is capable of being maintained in a plurality of strains. Corynebacterium-E. coli shuttle vectors include both an origin of replication of Corynebacterium and an origin of replication of E. coli. Use of shuttle vectors allows for easy introduction of a desired trait into a subject strain. For example, desired traits can be induced by cloning exogenous genes or mutation-inducing genes into a shuttle vector in E. coli, and then introducing the shuttle vector into Corynebacterium. 
There is a need for shuttle vectors which are able to maintain and proliferate in both Corynebacterium and E. coli, and which allow for easy accomplishment of a variety of cloning by improving a multi-cloning site.